EzCatDB D00537

EzCatDB: D00537

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DB code	D00537
RLCP classification	1.30.300.2 : Hydrolysis
CATH domain	Domain 1	2.-.-.-
CATH domain	Domain 2	3.20.20.40 : TIM Barrel		Catalytic domain
E.C.	3.2.1.91
CSA	1qk2

CATH domain	Related DB codes (homologues)
3.20.20.40 : TIM Barrel	S00194,D00536

Enzyme Name
UniProtKB			KEGG
	P07987	Q9C1S9	KEGG
Protein name	Exoglucanase 2	Exoglucanase-6A	cellulose 1,4-beta-cellobiosidase exo-cellobiohydrolase beta-1,4-glucan cellobiohydrolase beta-1,4-glucan cellobiosylhydrolase 1,4-beta-glucan cellobiosidase exoglucanase avicelase CBH 1 C1 cellulase cellobiohydrolase I cellobiohydrolase exo-beta-1,4-glucan cellobiohydrolase 1,4-beta-D-glucan cellobiohydrolase cellobiosidase
Synonyms	EC 3.2.1.91 Exoglucanase II Exocellobiohydrolase II CBHII 1,4-beta-cellobiohydrolase	EC 3.2.1.91 Exocellobiohydrolase 6A 1,4-beta-cellobiohydrolase 6A Beta-glucancellobiohydrolase 6A Avicelase 2
Pfam	PF00734 (CBM_1) PF01341 (Glyco_hydro_6) [Graphical view]	PF00734 (CBM_1) PF01341 (Glyco_hydro_6) [Graphical view]
CAZy	GH6 (Glycoside Hydrolase Family)	GH6 (Glycoside Hydrolase Family)

KEGG pathways
MAP code	Pathways
MAP00500	Starch and sucrose metabolism

UniProtKB:Accession Number	P07987	Q9C1S9
Entry name	GUX2_TRIRE	GUX6_HUMIN
Activity	Hydrolysis of 1,4-beta-D-glucosidic linkages in cellulose and cellotetraose, releasing cellobiose from the non- reducing ends of the chains.	Hydrolysis of 1,4-beta-D-glucosidic linkages in cellulose and cellotetraose, releasing cellobiose from the non- reducing ends of the chains.
Subunit		Monomer.
Subcellular location	Secreted.
Cofactor

Compound table: links to PDB-related databases & PoSSuM
								Substrates			Products
KEGG-id								C00760	C02013	C00001	C00185	C00760
Compound								Cellulose	Cellotetraose	H2O	Cellobiose	Cellulose
Type								polysaccharide	polysaccharide	H2O	polysaccharide	polysaccharide
ChEBI									62974	15377	17057
PubChem									439626	962 22247451	439178

1cb2A								Unbound	Unbound		Unbound	Unbound
1cb2B								Unbound	Unbound		Unbound	Unbound
1qjwA								Unbound	Analogue:GLC-GLC-SGC-MGL		Unbound	Unbound
1qjwB								Unbound	Analogue:GLC-GLC-SGC-MGL		Unbound	Unbound
1qk0A								Unbound	Analogue:GLC-XYS-IOB		Unbound	Unbound
1qk0B								Unbound	Analogue:GLC-XYS-IOB		Unbound	Unbound
1qk2A								Unbound	Analogue:GLC-GLC-SGC-MGL		Unbound	Unbound
1qk2B								Unbound	Analogue:GLC-GLC-SGC-MGL		Unbound	Unbound
1bvwA								Unbound	Unbound		Unbound	Unbound
2bvwA								Unbound	Unbound	Bound:HOH 97	Analogue:CTT	Analogue:GLC 602
2bvwB								Unbound	Unbound	Bound:HOH 431	Analogue:CT3	Analogue:GLC 603

Active-site residues
resource
literature [15]
pdb								Catalytic residues	comment

1cb2A								`ASP 175;ASP 221;ASP 263;ASP 401`	`mutant Y169F`
1cb2B								`ASP 175;ASP 221;ASP 263;ASP 401`	`mutant Y169F`
1qjwA								`ASP 175;ASP 221;ASP 263;ASP 401`	`mutant Y169F`
1qjwB								`ASP 175;ASP 221;ASP 263;ASP 401`	`mutant Y169F`
1qk0A								`ASP 175;ASP 221;ASP 263;ASP 401`
1qk0B								`ASP 175;ASP 221;ASP 263;ASP 401`
1qk2A								`ASP 175;ASP 221;ASP 263;ASP 401`
1qk2B								`ASP 175;ASP 221;ASP 263;ASP 401`
1bvwA								`ASP 180;ASP 226;ASP 268;ASP 405`
2bvwA								`ASP 180;ASP 226;ASP 268;ASP 405`
2bvwB								`ASP 180;ASP 226;ASP 268;ASP 405`

References for Catalytic Mechanism
References	Sections	No. of steps in catalysis
[14]	Scheme 1	1
[15]	Fig.6, Fig.7, p.8888-8890	1

references
[1]
Comments	X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
Medline ID	90333255
PubMed ID	2377893
Journal	Science
Year	1990
Volume	249
Pages	380-6
Authors	Rouvinen J, Bergfors T, Teeri T, Knowles JK, Jones TA
Title	Three-dimensional structure of cellobiohydrolase II from Trichoderma reesei.
Related UniProtKB	P07987
[2]
PubMed ID	1761039
Journal	Eur J Biochem
Year	1991
Volume	202
Pages	367-77
Authors	Gilkes NR, Claeyssens M, Aebersold R, Henrissat B, Meinke A, Morrison HD, Kilburn DG, Warren RA, Miller RC Jr
Title	Structural and functional relationships in two families of beta-1,4-glycanases.
[3]
PubMed ID	8399160
Journal	Biochemistry
Year	1993
Volume	32
Pages	9906-16
Authors	Spezio M, Wilson DB, Karplus PA
Title	Crystal structure of the catalytic domain of a thermophilic endocellulase.
[4]
PubMed ID	8192865
Journal	Biotechnol Appl Biochem
Year	1994
Volume	19
Pages	141-53
Authors	Woodward J, Brown JP, Evans BR, Affholter KA
Title	Papain digestion of crude Trichoderma reesei cellulase: purification and properties of cellobiohydrolase I and II core proteins.
[5]
PubMed ID	7733957
Journal	Biochem Biophys Res Commun
Year	1995
Volume	209
Pages	1046-52
Authors	Shultz MD, Lassig JP, Gooch MG, Evans BR, Woodward J
Title	Palladium--a new inhibitor of cellulase activities.
[6]
PubMed ID	7857933
Journal	Biochemistry
Year	1995
Volume	34
Pages	2220-4
Authors	Damude HG, Withers SG, Kilburn DG, Miller RC Jr, Warren RA
Title	Site-directed mutation of the putative catalytic residues of endoglucanase CenA from Cellulomonas fimi.
[7]
PubMed ID	8615816
Journal	Biochem J
Year	1996
Volume	315
Pages	467-72
Authors	Damude HG, Ferro V, Withers SG, Warren RA
Title	Substrate specificity of endoglucanase A from Cellulomonas fimi: fundamental differences between endoglucanases and exoglucanases from family 6.
[8]
PubMed ID	8856058
Journal	Eur J Biochem
Year	1996
Volume	240
Pages	584-91
Authors	Harjunpaa V, Teleman A, Koivula A, Ruohonen L, Teeri TT, Teleman O, Drakenberg T
Title	Cello-oligosaccharide hydrolysis by cellobiohydrolase II from Trichoderma reesei. Association and rate constants derived from an analysis of progress curves.
[9]
PubMed ID	9010760
Journal	J Biochem (Tokyo)
Year	1996
Volume	120
Pages	1123-9
Authors	Amano Y, Shiroishi M, Nisizawa K, Hoshino E, Kanda T
Title	Fine substrate specificities of four exo-type cellulases produced by Aspergillus niger, Trichoderma reesei, and Irpex lacteus on (1-->3), (1-->4)-beta-D-glucans and xyloglucan.
[10]
Comments	X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
Medline ID	97029636
PubMed ID	8875646
Journal	Protein Eng
Year	1996
Volume	9
Pages	691-9
Authors	Koivula A, Reinikainen T, Ruohonen L, Valkeajarvi A, Claeyssens M, Teleman O, Kleywegt GJ, Szardenings M, Rouvinen J, Jones TA, Teeri TT
Title	The active site of Trichoderma reesei cellobiohydrolase II: the role of tyrosine 169.
Related PDB	1cb2
Related UniProtKB	P07987
[11]
PubMed ID	8959766
Journal	Protein Expr Purif
Year	1996
Volume	8
Pages	399-400
Authors	Koivula A, Lappalainen A, Virtanen S, Mantyla AL, Suominen P, Teeri TT
Title	Immunoaffinity chromatographic purification of cellobiohydrolase II mutants from recombinant trichoderma reesei strains devoid of major endoglucanase genes.
[12]
PubMed ID	9204518
Journal	Appl Biochem Biotechnol
Year	1997
Volume	66
Pages	39-56
Authors	Medve J, Stahlberg J, Tjerneld F
Title	Isotherms for adsorption of cellobiohydrolase I and II from Trichoderma reesei on microcrystalline cellulose.
[13]
Comments	X-ray crystallography
PubMed ID	9662445
Journal	FEBS Lett
Year	1998
Volume	429
Pages	341-6
Authors	Koivula A, Kinnari T, Harjunpaa V, Ruohonen L, Teleman A, Drakenberg T, Rouvinen J, Jones TA, Teeri TT
Title	Tryptophan 272: an essential determinant of crystalline cellulose degradation by Trichoderma reesei cellobiohydrolase Cel6A.
Related PDB	1bvw
[14]
PubMed ID	9882628
Journal	Biochem J
Year	1999
Volume	337
Pages	297-304
Authors	Varrot A, Hastrup S, Schulein M, Davies GJ
Title	Crystal structure of the catalytic core domain of the family 6 cellobiohydrolase II, Cel6A, from Humicola insolens, at 1.92 A resolution.
[15]
Comments	X-ray crystallography
PubMed ID	10413461
Journal	Biochemistry
Year	1999
Volume	38
Pages	8884-91
Authors	Varrot A, Schulein M, Davies GJ
Title	Structural changes of the active site tunnel of Humicola insolens cellobiohydrolase, Cel6A, upon oligosaccharide binding.
Related PDB	2bvw
[16]
PubMed ID	10411622
Journal	Eur J Biochem
Year	1999
Volume	262
Pages	637-43
Authors	Carrard G, Linder M
Title	Widely different off rates of two closely related cellulose-binding domains from Trichoderma reesei.
[17]
Comments	X-ray crystallography
PubMed ID	10508787
Journal	Structure Fold Des
Year	1999
Volume	7
Pages	1035-45
Authors	Zou J, Kleywegt GJ, Stahlberg J, Driguez H, Nerinckx W, Claeyssens M, Koivula A, Teeri TT, Jones TA
Title	Crystallographic evidence for substrate ring distortion and protein conformational changes during catalysis in cellobiohydrolase Ce16A from trichoderma reesei.
Related PDB	1qk0,1qk2,1qjw
[18]
PubMed ID	10794732
Journal	Biochem J
Year	2000
Volume	348 Pt 1
Pages	201-7
Authors	Davies GJ, Brzozowski AM, Dauter M, Varrot A, Schulein M
Title	Structure and function of Humicola insolens family 6 cellulases: structure of the endoglucanase, Cel6B, at 1.6 A resolution.
[19]
PubMed ID	10601873
Journal	Eur J Biochem
Year	2000
Volume	267
Pages	244-52
Authors	Zhang S, Barr BK, Wilson DB
Title	Effects of noncatalytic residue mutations on substrate specificity and ligand binding of Thermobifida fusca endocellulase cel6A.
[20]
PubMed ID	11179652
Journal	FEMS Microbiol Lett
Year	2001
Volume	195
Pages	197-204
Authors	Lehtio J, Wernerus H, Samuelson P, Teeri TT, Stahl S
Title	Directed immobilization of recombinant staphylococci on cotton fibers by functional display of a fungal cellulose-binding domain.
[21]
PubMed ID	11325554
Journal	FEMS Microbiol Lett
Year	2001
Volume	198
Pages	57-63
Authors	Limon MC, Margolles-Clark E, Benitez T, Penttila M
Title	Addition of substrate-binding domains increases substrate-binding capacity and specific activity of a chitinase from Trichoderma harzianum.
[22]
PubMed ID	12007616
Journal	Biochim Biophys Acta
Year	2002
Volume	1596
Pages	366-80
Authors	Tuohy MG, Walsh DJ, Murray PG, Claeyssens M, Cuffe MM, Savage AV, Coughlan MP
Title	Kinetic parameters and mode of action of the cellobiohydrolases produced by Talaromyces emersonii.
[23]
PubMed ID	12188666
Journal	J Am Chem Soc
Year	2002
Volume	124
Pages	10015-24
Authors	Koivula A, Ruohonen L, Wohlfahrt G, Reinikainen T, Teeri TT, Piens K, Claeyssens M, Weber M, Vasella A, Becker D, Sinnott ML, Zou JY, Kleywegt GJ, Szardenings M, Stahlberg J, Jones TA
Title	The active site of cellobiohydrolase Cel6A from Trichoderma reesei: the roles of aspartic acids D221 and D175.
[24]
PubMed ID	12565856
Journal	Biochem Biophys Res Commun
Year	2003
Volume	301
Pages	280-6
Authors	Murray PG, Collins CM, Grassick A, Tuohy MG
Title	Molecular cloning, transcriptional, and expression analysis of the first cellulase gene (cbh2), encoding cellobiohydrolase II, from the moderately thermophilic fungus Talaromyces emersonii and structure prediction of the gene product.

comments

This family belongs to Glycosidase family-6, which has an inverting mechanism (equatorial to axial conformation). Furthermore, this enzyme belongs to exoglucanase, which requrires polymer-chain ends for catalytic activity, whilst its family member (EC 3.2.1.4) is endoglucanase.
The literature [7] suggests that endoglucanases from this family-6 may function by different mechanisms from this exoglucanase from the same family.
The literature [15] confirmed that Asp226 (2bvw) atcs as proton donor (acid catalyst) and Asp405 as the catalytic base. In sharp contrast, the literature [6] doubted the role of Asp405, which suggests Asp268 might act as base catalyst. In any case, the identification of catalytic base is difficult for these enzymes.

created	updated
2002-10-15	2009-09-29

Copyright: Nozomi Nagano, JST & CBRC-AIST
Funded by PRESTO/Japan Science and Technology Corporation (JST) (December 2001 - November 2004)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
Funded by Grant-in-Aid for Scientific Research (B)/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (September 2005 - September 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (October 2007 - September 2010)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2011 - March 2012)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2012 - March 2013)
Supported by the commission for the Development of Artificial Gene Synthesis Technology for Creating Innovative Biomaterial from the Ministry of Economy, Trade and Industry (METI) (October 2012 - )

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